Cholesterol e test wako kit

The average particle size and zeta-potential of liposomes were measured as described previously [11 (link)]. The lipid concentrations of Liposome-daunorubicin and Fuc-Liposome-daunorubicin were measured using a Cholesterol E-test Wako Kit as described previously [11 (link)].

The concentrations of cholesterol, bile acids, phospholipids and TG, and the activity of alanine aminotransferase (ALT) in each sample were measured using commercially available kits according to manufacturer’s instructions. The Cholesterol E-test Wako Kit, the TBA test Wako Kit (for the determination of total bile acids), the Phospholipids C-test Wako Kit, the Triglyceride E-test Wako Kit, and the Transaminase CII-test Wako Kit (Wako Pure Chemical Industries, Tokyo, Japan) were used in this study.

DXR was encapsulated into liposomes by a remote loading method [16 (link)]. The thin lipid film prepared as described above was hydrated with 300 mM citric acid solution and then vesiculated by sonication. The liposomes were passed through a Sephadex G-50 column equilibrated with 10% sucrose and then the concentration of cholesterol in liposomes was determined by Wako Cholesterol E test kit (Wako, Oosaka, Japan). DXR solution was mixed with liposomes at a concentration of 0.2 mg/mg lipid, followed by incubation at 65 °C for 1 h. DXR-encapsulated liposomes were purified through Sephadex G-50 columns equilibrated with PBS. To determine the concentration of DXR in liposomes, the optical absorbance (490 nm) was measured following the disruption of liposomes with 1% Triton-X. To investigate the intracellular delivery of DXR, A375 cells (5 × 10³ cells) were seeded into 96-well imaging plates coated with PLL. After 24 h cultivation, cells were treated with DXR-encapsulated liposomes at 0.5 μM DXR concentration at 37 °C for 6 h. DXR taken up by cells was observed by CLSM. The ratio of DXR-positive cells was estimated from five individual images.